Development of a luciferase/luciferin cell proliferation (XenoLuc) assay for real-time measurements of Gfp-Luc2-modified cells in a co-culture system

BMC Biotechnol. 2019 Jun 14;19(1):34. doi: 10.1186/s12896-019-0528-4.

Abstract

Background: In vitro modelling of cancer cells is becoming more complex due to prevailing evidence of intimate interactions between cancer cells and their surrounding stroma. A co-culture system which consists of more than one cell type is physiologically more relevant and thus, could serve as a useful model for various biological studies. An assay that specifically detects the phenotypic changes of cancer cells in a multi-cellular system is lacking for nasopharyngeal carcinoma (NPC).

Results: Here, we describe a luciferase/luciferin (XenoLuc) assay that could specifically measure changes in the proliferation of cancer cells in the co-culture system using two modified NPC patient-derived tumour xenograft (PDTXs) cells: Xeno284-gfp-luc2 and XenoB110-gfp-luc2. Through this assay, we are able to show that the growth of NPC xenograft cells in both two-dimensional (2D) and three-dimensional (3D) models was enhanced when co-cultured with normal human dermal fibroblasts (NHDFs). In addition, potential applications of this assay in in vitro drug or inhibitor screening experiments are also illustrated.

Conclusions: XenoLuc assay is specific, sensitive, rapid and cost-effective for measuring the growth of luciferase-expressing cells in a co- or multiple-culture system. This assay may also be adapted for tumour microenvironment studies as well as drug screening experiments in more complex 3D co-culture systems.

Keywords: Cancer immunology; Co-culture; Drug screening; Fibroblasts; Luciferase assay; Luminescence; Nasopharyngeal carcinoma; Nasopharyngeal neoplasms; Patient-derived xenograft; Proliferation; Spheroids; Stromal interaction; Tumour microenvironment.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Cells, Cultured
  • Coculture Techniques
  • Fibroblasts / cytology
  • Fibroblasts / metabolism*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism*
  • HEK293 Cells
  • Humans
  • Luciferases / genetics
  • Luciferases / metabolism*
  • Luminescent Measurements / methods
  • Mice, Inbred NOD
  • Mice, Knockout
  • Mice, SCID
  • Nasopharyngeal Carcinoma / genetics
  • Nasopharyngeal Carcinoma / metabolism*
  • Nasopharyngeal Carcinoma / pathology
  • Transplantation, Heterologous

Substances

  • Green Fluorescent Proteins
  • Luciferases